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Published December 2023 | Published
Journal Article

Assembly of complete mouse embryo models from embryonic and induced stem cell types in vitro

  • 1. ROR icon California Institute of Technology


The interaction between embryonic and extraembryonic tissues is critical in natural mouse embryogenesis. Here, to enable such interaction in vitro, we describe a protocol to assemble a complete mouse embryo model using mouse embryonic stem cells and induced embryonic stem cells to express Cdx2 (or trophoblast stem cells) and Gata4 to reconstitute the epiblast, extraembryonic ectoderm and visceral endoderm lineages, respectively. The resulting complete embryo models recapitulate development from embryonic day 5.0 to 8.5, generating advanced embryonic and extraembryonic tissues that develop through gastrulation to initiate organogenesis to form a head and a beating heart structure as well as a yolk sac and chorion. Once the required stem cell lines are stably maintained in culture, the protocol requires 1 day to assemble complete embryo models and a further 8 days to culture them until headfold stages, although structures can be collected at earlier developmental stages as required. This protocol can be easily performed by researchers with experience in mouse stem cell culture, although they will benefit from knowledge of natural mouse embryos at early postimplantation stages.

Copyright and License

© Crown 2023.


The authors thank S. Raj for providing the images for Fig. 3, K. Ivanovitch for advice on the handling and storage of rat serum and M.Z.-G. laboratory members for their helpful comments. This work was supported by National Institutes of Health Pioneer Award (DP1 HD104575-01), the Allen Discovery Center for Lineage Tracing; European Research Council (669198), the Wellcome Trust (207415/Z/17/Z), Open Philanthropy/Silicon Valley Community Foundation and Weston Havens Foundation. K.Y.C.L. was supported by the Croucher Foundation and the Cambridge Trust.


These authors contributed equally: Kasey Y. C. Lau, Gianluca Amadei.

K.Y.C.L. performed the experiments. K.Y.C.L. and G.A. developed the protocol with the guidance from M.Z.-G. M.Z.-G. conceived and supervised the project. K.Y.C.L., G.A. and M.Z.-G. wrote the manuscript.

Data Availability

Supporting data of this study can be found in our previous publications7,8,13 and are available from the corresponding author upon request.

Conflict of Interest

A patent 'Generation of synthetic embryos from multiple stem cell types' was filed by California Institute of Technology and the University of Cambridge under CIT file number: CIT-8826-P and serial number: 63/344,251.

Additional details

October 12, 2023
January 9, 2024